Identification of the arginine/ornithine antiporter ArcD from Halobacterium salinarum.
This paper identifies the first arginine/ornithine antiporter ArcD from the domain of archea. The functional role of ArcD is demonstrated by transport assays with radioactive labelled arginine, by its necessity to enable arginine fermentation under anaerobic growth conditions and by the consumption of arginine from the medium during growth. All three experimentally observables are severely disturbed when the deletion strain DeltaArcD is used. The isolated protein is verified by mass spectrometry and reconstituted in vesicles. The proteoliposomes are attached to a membrane and capacitive currents are recorded which appear upon initiation of the transport process by change from arginine-free to arginine-containing buffer. This clearly demonstrates that the purified 34kD protein is the functional unit.
Wimmer
Florian
F
Department of Membrane Biochemistry, Max Planck Institut for Biochemistry, Am Klopferspitz 18, D82152 Martinsried, Germany.
Oberwinkler
Tanja
T
Bisle
Birgit
B
Tittor
Jörg
J
Oesterhelt
Dieter
D
D000595:Amino Acid Sequence; D017920:Antiporters / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001120:Arginine / Q000378:metabolism*; D001692:Biological Transport; D019617:Halobacterium salinarum / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009952:Ornithine / Q000378:metabolism*; D017433:Protein Structure, Secondary
D017920:Antiporters; D019843:Archaeal Proteins; D001120:Arginine; D009952:Ornithine